Department of Applied Chemistry and Biochemistry, Graduate School of Science and Technology, Kumamoto University, Chuo-ku, Kumamoto 860-8555, Japan; Japan Science and Technology (JST), Core Research for Evolutional Science and Technology (CREST), Chiyoda-ku, Tokyo 102-0075, Japan.
Anal Biochem. 2013 Dec 1;443(1):41-5. doi: 10.1016/j.ab.2013.08.010. Epub 2013 Aug 19.
Copolymer particles for removal of endotoxins (lipopolysaccharides, LPSs) were prepared by suspension copolymerization of γ-cyclodextrin (CyD) and 1,6-hexamethylenediisocyanate. The LPS-removing activity of the copolymer particles was compared with that of poly(ε-lysine)-immobilized Cellufine (cationic adsorbent) or polystyrene particles (hydrophobic adsorbent) by a batch method. When DNA was present in solution with LPSs under physiological conditions (pH 6.0, ionic strength of μ = 0.05-0.8), LPS-removing activity of the cationic or hydrophobic adsorbent was unsatisfactory because both the DNA and the LPSs were adsorbed onto each adsorbent. By contrast, the copolymer particles with γ-CyD cavity (CyD content: 14-20 mol%) could selectively remove LPSs from a DNA solution (50 μg ml(-1), pH 6.0, and μ = 0.05-0.2) containing LPSs (15 EU ml(-1)) without the adsorption of DNA. The residual concentration of LPSs in the treated DNA solution was below 0.1 EU ml(-1), and the recovery of DNA was 99%.
用于去除内毒素(脂多糖,LPS)的共聚物颗粒通过 γ-环糊精(CyD)和 1,6-己二异氰酸酯的悬浮共聚制备。通过分批法将共聚物颗粒的 LPS 去除活性与聚(ε-赖氨酸)固定化 Cellufine(阳离子吸附剂)或聚苯乙烯颗粒(疏水性吸附剂)进行比较。当 DNA 与 LPS 在生理条件下(pH 6.0,μ = 0.05-0.8)存在于溶液中时,阳离子或疏水性吸附剂的 LPS 去除活性并不令人满意,因为 DNA 和 LPS 都被吸附到每个吸附剂上。相比之下,具有 γ-CyD 腔的共聚物颗粒(CyD 含量:14-20 mol%)可以从含有 LPS(15 EU ml(-1))的 DNA 溶液(50 μg ml(-1),pH 6.0,μ = 0.05-0.2)中选择性地去除 LPS,而不会吸附 DNA。处理后的 DNA 溶液中 LPS 的残留浓度低于 0.1 EU ml(-1),并且 DNA 的回收率为 99%。